Design and In vitro Evaluation of Branched Peptide Conjugates: Turning Nonspecific Cytotoxic Drugs into Tumor‐Selective Agents

The use of peptide receptors as targets for tumor‐selective therapies was envisaged years ago with the findings that receptors for different endogenous regulatory peptides are overexpressed in several primary and metastatic human tumors, and can be used as tumor antigens. Branched peptides can retain or even increase, through multivalent binding, the biological activity of a peptide and are very resistant to proteolysis, thus having a markedly higher in vivo activity compared with the corresponding monomeric peptides. Oligo‐branched peptides, containing the human regulatory peptide neurotensin (NT) sequence, have been used as tumor‐specific targeting agents. These peptides are able to selectively and specifically deliver effector units, for cell imaging or killing, to tumor cells that overexpress NT receptors. Results obtained with branched NT conjugated to different functional units for tumor imaging and therapy indicate that branched peptides are promising novel multifunctional targeting molecules. This study is focused on the role of the releasing pattern of drug‐conjugated branched NT peptides. We present results obtained with oligo‐branched neurotensin peptides conjugated to 6‐mercaptopurin (6‐MP), combretastain A‐4 (CA4) and monastrol (MON). Drugs were conjugated to oligo‐branched neurotensin through different linkers, and the mode‐of‐release, together with cytotoxicity, was studied in different human cancer cell lines. The results show that branched peptides are very promising pharmacodelivery options. Among our drug‐armed branched peptides, NT4–CA4 was identified as a candidate for further development and evaluation in preclinical pharmacokinetic and pharmacodynamic studies. This peptide–drug exhibits significant activity against pancreas and prostate human cancer cells. Consequently, this derivative is of considerable interest due to the high mortality rates of pancreas neuroendocrine tumors and the high incidence of prostate cancer.     

Differential scanning calorimetry of lupin and soy proteins

Differential scanning calorimetry (DSC) was used to study the 7S and 11S globulin fractions extracted from lupin seed (Lupinus luteus) flour. In agreement with previous work on other lupin species, the isolate showed three denaturation peaks compared to the two observed with soy. By comparison with the isolated globulin fractions, the denaturation peaks at the two higher temperatures in the lupin isolate were assigned to the 11S and 7S globulins. The denaturation temperature of the lupin 7S globulin was about 10 K higher than that for the corresponding soy globulin, whereas the values for the 11S globulin were similar. All globulins displayed increasing thermal stability with decreasing moisture contents. Possible reasons for the differences in behaviour of soy and lupin protein isolates are discussed.     

Detection of the adulteration of olive oils by solid phase microextraction and multidimensional gas chromatography

The presence or absence of filbertone in 21 admixtures of olive oil with virgin and refined hazelnut oils obtained using various processing techniques from different varieties and geographical origins was evaluated by solid phase microextraction and multidimensional gas chromatography (SPME–MDGC). The obtained results showed that the sensitivity achievable with the proposed procedure was enough to detect filbertone and, hence, to establish the adulteration of olive oil of different varieties with virgin hazelnut oils in percentages of up to 7%. The very low concentrations in which filbertone occurs in some refined hazelnut oils made difficult its detection in specific admixtures. In any case, the minimum adulteration level to be detected depends on the oil varieties present in the adulterated samples. In the present study, the presence of R- and S-enantiomers of filbertone could be occasionally detected in olive oils adulterated with 10–20% of refined hazelnut oil.     

Evaluation of the spiral plating method for the enumeration of microorganisms throughout the manufacturing and ripening of a raw goat's milk cheese

A statistical comparison of the spiral plate count (SPLPC) and the standard plate count (SPC) methods for enumeration of microorganisms in raw goat's milk cheese throughout its manufacturing and ripening was carried out. Enumeration of mesophiles, lactic acid bacteria (presumptive lactococci, presumptive leuconostocs, and presumptive lactobacilli), Micrococcaceae, Enterobacteriaceae, and molds and yeasts was carried out for milk, curd, and 2-, 5-, 10-, 17-, and 27-day-old cheeses. Average counts for the SPLPC and SPC methods differed by less than half of a log cycle for all microbial groups studied (range of difference, -0.1386 [mesophiles] to +0.4397 [presumptive lactobacilli]). The results of the SPLPC method compared favorably with the results of the SPC procedure for mesophiles, presumptive lactococci, presumptive leuconostocs, Enterobacteriaceae, and molds and yeasts (the variance between replicate platings was close to 0.005, and correlation coefficients were >0.9). Correlation coefficients were lower for Micrococcaceae (r = 0.824) and presumptive lactobacilli (r = 0.670). Analysis of variance showed that the plating method was a significant factor (P < 0.05) for presumptive lactobacilli counts. In general, results from the SPLPC method compared favorably with results from SPC procedure in the enumeration of microorganisms in goat cheese throughout its manufacturingand ripening processes. However, the suitability of the SPLPC method depends mainly on the microbial group studied.     

Hot water and organic acid interventions to control microbiological contamination on hog carcasses during processing

Pork skin and muscle tissue were washed with water at temperatures from 25 to 80 degrees C. Water temperatures of 65 and 80 degrees C resulted in greater population reductions of Enterobacteriaceae on pork muscle tissue than lower water temperatures. There was no observable effect of water temperature on population reductions of Enterobacteriaceae on pork skin. Water temperatures of 55, 65, and 80 degrees C reduced the populations of Enterobacteriaceae on inoculated scalded carcasses processed in a university abattoir by 1 to 1.5 log/cm2. Following the water wash with an organic acid rinse resulted in further numerical reductions in populations, although these were not statistically different from the water wash alone. The jowls of both scalded and skinned carcasses processed in a commercial establishment were directly inoculated with a fecal material slurry and then processed with organic acid rinsing only, hot water washing only, or a combination of hot water washing followed by organic acid rinsing. The hot water and acid treatment reduced the populations of mesophilic aerobic bacteria and Escherichia coli by approximately 2 log cycles on both scalded and skinned hog carcasses. The combined treatment resulted in 60% of the scalded carcasses and 40% of the skinned carcasses with undetectable levels of E. coli after direct fecal inoculation of the carcasses. Hot water washing followed by organic acid rinsing can significantly improve the microbiological quality of pork carcasses.     

A log square average case algorithm to make insertions in fast similarity search

To speed up similarity based searches many indexing techniques have been proposed in order to address the problem of efficiency. However, most of the proposed techniques do not admit fast insertion of new elements once the index is built. The main effect is that changes in the environment are very costly to be taken into account.In this work, we propose a new technique to allow fast insertions of elements in a family of static tree-based indexes. Unlike other techniques, the resulting index is exactly equal to the index that would be obtained by building it from scratch. Therefore there is no performance degradation in search time.We show that the expected number of distance computations (and the average time complexity) is bounded by a function that grows with log²(n) where n is the size of the database.In order to check the correctness of our approach some experiments with artificial and real data are carried out.     

A neural network based intelligent system for tile prefetching in web map services

Web mapping has become a popular way of distributing interactive digital maps over the internet. Instead of dynamically generating map images on the fly, those can be pre-generated and served from a server-side cache for faster retrieval. However, these caches can grow unmanageably in size when the cartography covers mid to large areas for multiple rendering scales. This forces modest organizations to use partial caches containing just a subset of the total tiles, and makes their services less attractive than other mapping services like Google Maps or Microsoft Bing Maps. This work proposes a neural-network-based intelligent system that predicts which areas are likely to be requested in the future from a catalog of geographic features and a short history of past requests. These priority regions can be used by a tile prefetching policy to achieve an optimal population of the cache. Neural networks are trained and validated using supervised learning with real data-sets from a public nation-wide web map service. Trace-driven simulations demonstrate that accurate long-term predictions, up to 90% in terms of cache-hit ratio, can be obtained with the proposed model by prefetching a low fraction, only the 20% of the total tiles, and with a short training period.